Jemds.

com Original Research Article

A COMPARATIVE STUDY OF ABO BLOOD GROUPS AND SECRETOR STATUS IN ISCHAEMIC HEART
DISEASE PATIENTS IN KADAPA CITY
Parveen Shaik1, A. Chandra Sekhar2

1AssistantProfessor, Department of Physiology, RIMS, Kadapa, Andhra Pradesh, India.

2AssociateProfessor and HOD I/C, Department of Physiology, RIMS, Kadapa, Andhra Pradesh, India.
ABSTRACT
BACKGROUND
Obesity, hypertension, diabetes mellitus, dyslipidaemia, smoking, stress, sedentary lifestyle and genetic factors are some well-
known risk factors for Ischaemic Heart Disease (IHD). The ABO blood groups and secretor status may be linked to IHD and its risk
factors.
Aim- To study the correlation between ABO blood groups and secretor status in the IHD patients in the city of Kadapa.

MATERIALS AND METHODS

This study was conducted in Department of Physiology, RIMS Medical College, from August 2015 to January 2016. This is a case
control study and selection of cases and controls was done by simple random sampling. 600 patients with IHD diagnosed based on
the electrocardiograms were chosen from Cardiology Unit, RIMS General Hospital, Kadapa. 600 volunteer blood donor’s age and
sex matched with no evidence of any disease were included. Blood group estimation was done by slide agglutination method and
secretor status was done by indirect haemagglutination method. The sample size required was taken for convenience. Chi-square
test and p-value were used for statistical analysis by SPSS S
oftware version 17.

RESULTS
The frequency distribution among IHD patients was maximum in B group (45.6 %) followed by O group (21.6%), A group (19.9%)
and AB group (12.7%). Among the cases, 21.5% were secretors and 78.5% were non-secretors and there was a significant
association of non-secretors with IHD with Chi-square value of 401.3 and a p-value of 0.00000.

CONCLUSION
The results of this study conclude that B group non-secretors are more prone to IHD in the city of Kadapa.

KEY WORDS
ABO Blood Groups, Secretor Status, Ischaemic Heart Disease.
HOW TO CITE THIS ARTICLE: Shaik P, Sekhar AC. A comparative study of ABO blood groups and secretor status in ischaemic
heart disease patients in Kadapa city. J. Evolution Med. Dent. Sci. 2018;7(42):4545-4549, DOI: 10.14260/jemds/2018/1014
BACKGROUND
At the turn of 20th century, Karl Landsteiner first described
the existence of serological differences between individuals
and stated that people of the world, irrespective of their race
can be divided into four groups depending on the substances
present on the surface of their red blood cells. In 1901, he
grouped the individuals into A, B, AB and O. The discovery of
the iso-agglutinogens was a milestone in the field of medicine.
Karl Landsteiner received the Nobel prize for his discovery of
the ABO system of blood groups.(1)
The A, B and O genes all locate together at 9q34.1 - q34.2.
The genes of the ABO system do not encode directly for the
antigens, but encode for enzymes that add specific sugars to
the red cell membrane. These sugars are the ABO red cell
antigens that are detectable with serological testing. The A
gene codes for the transferase α(1,2)
‘Financial or Other Competing Interest’: None.
Submission 22-08-2018, Peer Review 27-09-2018,
Acceptance 03-10-2018, Published 15-10-2018.
Corresponding Author:
Dr. A. Chandra Sekhar,
Associate Professor,
Department of Physiology,
RIMS, Kadapa-516001,
Andhra Pradesh, India.
E-mail: sekharphysiology@gmail.com
DOI: 10.14260/jemds/2018/1014
N-acetylgalactosamine transferase and the B gene codes
for transferase α[1,2] galactosyl transferase and O allele
encodes for non-functional transferase.(3)
The ABH antigens are found not only on red cells, but also
on other cells in the most body fluids and on the cell
membranes of tissues such as intestine, urothelium and
vascular endothelium. The expression of ABH antigens into
body fluids is controlled by the Sese genes and they are
located on chromosome 19q13.3.(2)
There is some evidence that ABO blood groups may be
associated with certain diseases. Gastric cancer has been
reported to be more prevalent in individuals with blood
group A, but peptic ulcer is more often seen in those with
blood group O.(4)
The term ABH secretor refers to secretion of ABO blood
group antigens in fluids such as saliva, sweat, tears, semen
and serum. Approximately, 80% of people are secretors (SeSe
or Sese). People who do not secrete their blood type antigen
in their secretions are termed non-secretors. About 15% of
the population are non-secretors. For example-
 O Group secrete H antigens.
 A group secrete A and H antigens.
 B group secrete B and H antigens.
 AB group secrete A, B and H antigens.(5)
IHD is a condition in which there is an inadequate supply
of blood and oxygen to a portion of the myocardium. It

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typically occurs when there is an imbalance between
myocardial oxygen supply and demand.(6) IHD causes more
deaths and disability and incurs greater economic costs than
any other illness in the developed world. Risk factors that are
associated with IHD are high fat and energy rich diet,
smoking, sedentary lifestyle, obesity, insulin resistance and
type II diabetes mellitus.(6,7,8) The way to reduce Coronary
Artery Disease (CAD) risk include eating a healthy diet,
regular exercise, maintaining a healthy weight and not
smoking.(9)
MATERIALS AND METHODS
Study Design/ Method of Study
This is a case-control study, which is a type of observational
study.
Method of Sampling
Simple random sampling was used among cases and controls.
This study was conducted between August 2015 and
January 2016 in RIMS General Hospital, Kadapa, Andhra
Pradesh state, India. Case sheets were filled for the IHD
patients and control subjects to obtain their medical history
and socio-demographic parameters (Age, sex, educational
status, occupation, blood groups and willingness to
participate in the study). The sample size required was taken
for convenience.
Cases
The study included a total of 600 IHD patients. The IHD was
diagnosed based on ECG taken who came to Cardiology unit,
RIMS General Hospital, Kadapa complaining of chest pain.
Patients were selected at random after admission from ICCU
(Intensive Coronary Care Unit). Patients of both sexes were
selected in the age group of 25 - 65 years. 600 patients were
studied, among them 444 were males and 156 were females.
Controls
600 Control subjects were recruited for this study from the
healthy volunteer blood donors with no evidence of any
disease who came to blood bank, RIMS General Hospital,
Kadapa.
Both males and females of ages ranging from 25 to 65
years were recruited. Among 600 controls, 412 were males
and 188 were females.
Blood Group Determination
Blood group was determined by slide agglutination
technique.
Principle
The surface of the red cell membrane contains genetically
determined antigens called agglutinogens, while plasma
contains antibodies called agglutinins. To determine the
blood group of a person, his/ her red cells are made to react
with sera containing agglutinins. The slide is then examined
under a microscope to detect the presence or absence of
clumping and haemolysis of red cells that occurs as a result of
antigen-antibody reaction.
J. Evolution Med. Dent. Sci./eISSN- 2278-4802, pISSN- 2278-4748/ Vol. 7/ Issue 42/ Oct. 15, 2018
Original Research Article
Materials
Antisera, slide, lancet, compound microscope.
Procedure
Under aseptic precautions, the pulp of the ring finger was
pricked by a sterile lancet and one drop of anti-A was placed
on one side of a microscopic slide and labelled as A. One drop
of anti-B was placed on the other side of the same slide and
labelled as B. A drop of blood was added to each drop of
antiserum. Blood groups were determined as follows-
 Agglutination in slide A- Blood Group A
 Agglutination in slide B- Blood Group B
 Agglutination in both slides- Blood Group AB
 Agglutination in neither slide- Blood Group O
Determination of Secretor Status
It is determined by Haemagglutination inhibition technique.
Principle
If the blood group antigens are present in saliva when an
appropriate antiserum is added to it, an antigen-antibody
reaction occurs. The antibodies in the serum neutralise the
antigens in saliva. When a red cell suspension of the same
blood group is now added to this mixture, there is no
agglutination due to previous inhibition of the antiserum.
Thus, in the case of secretors there will be no agglutination
seen. In the case of non-secretors, as their saliva does not
contain blood group antigens, the antiserum added will not
be inhibited by the antigens. Now when the red cell
suspension of the same blood group is added to the mixture
and there will be an agglutination reaction. Thus, in case of
non-secretors, there will be agglutination.
Materials
1. 5% red cell suspension.
2. Antiserum (Anti-A, Anti-B, Anti-H)
3. Diluted and processed saliva.
4. Test tube rack with tubes.
5. Pipette.
6. Microscope.
7. Slide.
8. Centrifuge.
9. Hot water bath.
10. Sterile containers.
Procedure
Saliva was collected at room temperature between 11 a.m.
and 12 noon and tests were carried out between 12 noon and
4 p.m.
a) Preparation of 5% Red Cell Suspension
The freshly collected blood in an EDTA bottle was transferred
into a small glass tube and centrifuged at a speed of 3000 rpm
for 15 minutes to pack the red cells. The supernatant plasma
was separated as much as possible from the cells and
replaced by sterile isotonic saline. This mixture was
centrifuged and again the supernatant separated from the
cells. This procedure was repeated 3 to 4 times to wash the
red blood cells. Washing of the red blood cells was done to
remove any antibody present on the cells.
After the last washing, the supernatant was removed and
one drop of packed red cells was mixed with 19 drops of
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normal saline to prepare a 5% red cell suspension in isotonic
saline.

b) Collection and Processing of Saliva

Saliva was collected at room temperature in sterile
disposable containers. The patient was instructed to wash the
mouth and drink a glass of water. One millilitre of saliva was
collected in a sterile plastic bottle and transferred to a sterile
test tube. The tube was then kept in a boiling water bath
(100oc) for twenty minutes. Saliva was boiled to destroy the
enzymes present. The saliva was then centrifuged at 3000
revolutions per minute (rpm) for fifteen minutes. The
supernatant was separated and diluted in a 1: 4 ratio. All the
saliva samples were tested for secretor status on the same Blood Groups Males Females
day of their collection. A 90 30
B 184 92
c) Dilution of Anti-Serum
AB 60 18
Anti-A and Anti-B antisera were used for patients of Blood
O 110 16
Group A and Blood Group B and Anti-H antisera was used for
Table 1. Gender Wise distribution of Cases
patients belonging to Blood Group O. Antisera were diluted in
a dilution of 1: 8. E.g. if diluted anti-A was to be prepared,
Chi-square value 19.7
four test tubes were kept in a row and one drop of saline was
‘P’ value 0.00005
put into each tube. One drop of Anti-A was now added to the
first tube and mixed. Then one drop of diluted serum was
Blood Groups Males Females
taken from the first tube and added to the second tube. The
A 42 36
same procedure was followed up to the fourth tube. (Ref) as
B 150 42
shown in the table below:
AB 40 32
O 180 78
Tube Dilution
Table 2. Gender Wise distribution of Controls
Tube 1 1:1
Tube 2 1:2
Chi-square value 21.8439
Tube 3 1:4
‘P’ value 0.00000
Tube 4 1:8

One drop of processed saliva and one drop of diluted

antiserum from test tube four (1: 8 dilution) were now added
to fresh test tube.

d) Procedure to determine Secretor Status

The contents were mixed well and allowed to stand for fifteen
minutes. Then a drop of standard red cell suspension was
added and allowed to stand for sixty minutes. The mixed
solution was examined under microscope. Care was taken to
wash the pipette repeatedly, immediately after each dip. No
agglutination suggested the presence of antigen in saliva,
while agglutination suggested the absence of antigens in the
Blood Secretors Non-Secretors
saliva. Saline controls were kept simultaneously with the test.
A 26 94
RESULTS B 61 215
The study was done to find out that there is any correlation AB 18 60
between ABO blood groups, secretor status and the incidence O 24 102
of IHD in Kadapa district. 600 IHD patients (444 males and Table 3. Distribution of Secretor Status among Cases
156 females) and 600 control (412 males and 188 females)
subjects participated in the study. Chi-square value 0.6250
A total of 129 (21.5%) IHD patients and 476 (79.4%) ‘P’ value 0.8907
control subjects were secretors, while 471 (78.5%) IHD
patients and 124 (20.6%) control subjects were non-
secretors.

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DISCUSSION
The frequency of ABH secretors and non-secretors in the
Secretors Non-Secretors control subjects were 79.4% (476) and 20.6% (124)
respectively. Whereas, the frequencies of secretors and non-
A 63 15
secretors in the IHD patients were 21.5% and 78.5%
B 157 35
respectively. The frequency of ABH secretor status in the
AB 56 16
control subjects in this study is generally higher than what is
O 200 58 obtainable worldwide, where about 20.6% are non-
Table 4. Distribution of Secretors and Non-Secretors secretors.(10) Igbeneghu et al(11) reported a frequency of 75%
among Controls secretors and 25% non-secretors in Osogbo in south-western
Nigeria, while Jaff(12) reported a frequency of 76% secretors
Chi-square 1.4179 and 24% non-secretors in Iraq. Akhter et al(13) found a
‘P’ value 0.7013 frequency of ABH secretor status of 60% and non-secretors of
40% in Dhakar.
The FUT2 gene encodes fucosyltransferases that transfer
a terminal fucose residue to a pre-existing precursor
substance to form a soluble H antigen in secretory tissues,
which serves as a precursor for soluble ABH antigens. Hence,
individuals having at least one functional FUT2 allele, their
ABH antigens are not only detected on their cell surfaces, but
also in their body fluids including saliva. Non-secretors are
homozygous for two inactive FUT2 alleles (SeSe). (14) It is
therefore possible that the presence of genes that predispose
a person to IHD may down-regulate the expression of FUT2
(secretor) gene and secrete enzymes, leading to the higher
percentage of non-secretors in IHD patients.

CONCLUSION
The study concludes that there is a significant correlation
between ABO blood types and secretor status. The incidence
of IHD is more common in B group non-secretors in Kadapa
city. This is in correlation with study done by Meian He
et al(15) that non-O blood group had higher risk of Coronary
Heart Disease (CHD),(16,17,18,19) because factor VIII-vWF levels
are a risk for CHD.(20,21) IHD was associated with ‘A’ blood
Blood Groups Secretors Non-Secretors group in studies of Saima Sharif et al(22) and was associated
Cases 129 471 with ‘B’ group in studies of Shazia Chowdhary et al.(23)
Controls 476 124
Table 5. Distribution of Secretors and Non-Secretors Recommendations
among Cases and Controls To reduce the risk, lifestyle modifications like physical
exercise, antioxidant rich diet like fruits, vegetables and
Chi-square value 401.3 polyunsaturated fats contained at an early age by reducing
‘P’ value 0.00000 the role of risk factors for ischaemic heart disease as secretor
status and ABO types are genetically determined.(24)

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